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BD Horizon™ BV421 Mouse Anti-Human CD158f
Clone UP-R1 (RUO)
Multicolor flow cytometric analysis of CD158f expression on human peripheral blood CD3- lymphocytes. Human whole blood was stained with PE Mouse Anti-Human CD56 (Cat. No. 555516/561903) and APC Mouse Anti-Human CD3 (Cat. No. 555335/561810/561811) antibodies, and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Human CD158f antibody (Cat. No. 566330; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric contour plots showing the correlated expression of CD158f (or Ig Isotype control staining) versus CD56 were derived from CD3-negative gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Multicolor flow cytometric analysis of CD158f expression on human peripheral blood CD3- lymphocytes. Human whole blood was stained with PE Mouse Anti-Human CD56 (Cat. No. 555516/561903) and APC Mouse Anti-Human CD3 (Cat. No. 555335/561810/561811) antibodies, and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Human CD158f antibody (Cat. No. 566330; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric contour plots showing the correlated expression of CD158f (or Ig Isotype control staining) versus CD56 were derived from CD3-negative gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Multicolor flow cytometric analysis of CD158f expression on human peripheral blood CD3- lymphocytes. Human whole blood was stained with PE Mouse Anti-Human CD56 (Cat. No. 555516/561903) and APC Mouse Anti-Human CD3 (Cat. No. 555335/561810/561811) antibodies, and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Human CD158f antibody (Cat. No. 566330; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric contour plots showing the correlated expression of CD158f (or Ig Isotype control staining) versus CD56 were derived from CD3-negative gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Multicolor flow cytometric analysis of CD158f expression on human peripheral blood CD3- lymphocytes. Human whole blood was stained with PE Mouse Anti-Human CD56 (Cat. No. 555516/561903) and APC Mouse Anti-Human CD3 (Cat. No. 555335/561810/561811) antibodies, and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Human CD158f antibody (Cat. No. 566330; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric contour plots showing the correlated expression of CD158f (or Ig Isotype control staining) versus CD56 were derived from CD3-negative gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Product Details
BD Horizon™
CD158F; CD158F1; CD158F2; KI2LA; KIR2DL5; KIR2DL5A; KIR2DL5B
Human (QC Testing)
Mouse BALB/c IgG1
Human CD158f Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl/test
AB_2739690
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
Lysing Buffer RUO
Size
100 ML
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 ML
Cat No.
349202
BV421 Mouse IgG1, k Isotype Control RUO
Size
50 µg
Cat No.
562438
566330 Rev. 1
Antibody Details
UP-R1
The UP-R1 monoclonal antibody specifically recognizes CD158f which is also known as KIR2DL5. CD158f is a ~60 kDa single-pass type I transmembrane glycoprotein that belongs to the KIR (Killer cell Ig-like receptors) family within the Ig gene superfamily. CD158f is encoded by two gene paralogs, KIR2DL5A and KIR2DL5B that show greater than 99% identity in their coding sequences. CD158f molecules contain two extracellular C2-type Ig-like domains and a long cytoplasmic domain containing one canonical immunoreceptor tyrosine-based inhibitory motif (ITIM) and another atypical ITIM sequence. Although the exact ligands for CD158f have not yet been identified, the inhibitory function of UP-R1 antibody-crosslinked CD158f was shown to primarily depend upon a Src homology region 2-containing protein tyrosine phosphatase-2 (SHP-2) -dependent pathway. CD158f is highly polymorphic and is expressed on subsets of NK cells and T lymphocytes by some individuals. UP-R1 recognizes cell surface CD158f encoded by the most commonly expressed allele, KIR2DL5A*001. The UP-R1 antibody has been predicted to recognize CD158f molecules encoded by other transcriptionally-active KIR2DL5 alleles, A*012 and B*00602.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
566330 Rev. 1
Format Details
BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
407 nm
423 nm
566330 Rev.1
Citations & References
View product citations for antibody "566330" on CiteAb
Development References (5)
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Cisneros E, Moraru M, Gómez-Lozano N, López-Botet M, Vilches C. KIR2DL5: An Orphan Inhibitory Receptor Displaying Complex Patterns of Polymorphism and Expression.. Front Immunol. 2012; 3:289. (Clone-specific). View Reference
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Du Z, Sharma SK, Spellman S, Reed EF, Rajalingam R. KIR2DL5 alleles mark certain combination of activating KIR genes.. Genes Immun. 2008; 9(5):470-80. (Biology). View Reference
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Estefanía E, Flores R, Gómez-Lozano N, Aguilar H, López-Botet M, Vilches C. Human KIR2DL5 is an inhibitory receptor expressed on the surface of NK and T lymphocyte subsets.. J Immunol. 2007; 178(7):4402-10. (Immunogen: ELISA, Flow cytometry). View Reference
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Vilches C, Rajalingam R, Uhrberg M, Gardiner CM, Young NT, Parham P. KIR2DL5, a novel killer-cell receptor with a D0-D2 configuration of Ig-like domains.. J Immunol. 2000; 164(11):5797-804. (Biology). View Reference
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Yusa S, Catina TL, Campbell KS. KIR2DL5 can inhibit human NK cell activation via recruitment of Src homology region 2-containing protein tyrosine phosphatase-2 (SHP-2).. J Immunol. 2004; 172(12):7385-92. (Biology). View Reference
566330 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.